These are the continuing chronicles of prion disease, the reason for your own state of unease.
For past research and understanding of just what a prion is, see my previous research here:
I wish to pose a question…
Is it possible that, in our efforts to create synthetic drugs (for profit) in order to artificially mimic or replace the body’s natural health and healing processes, even while suppressing the body’s immune response to those drugs so that they may fool the natural system, we have inadvertently created a permanent state of dis-ease as the average human condition?
Let’s take an obscene example.
Over half a century ago, while researching the efficiency of the vaccine for smallpox, Japanese virologists working for the Institute For Infectious Diseases at University of Tokyo published their findings (1954) that some “viral inhibitory factor” was inhibiting the growth of their purposefully induced viral infection of laboratory research rabbits. In other words, the tiny rabbit bodies were having the natural immune response they should, which interferes with the capability of a foreign zoological pathogen to propagate (grow and reproduce) after injection. But they also discovered through isolation of this unknown and naturally occurring preventative substance that it was not originated from antibodies. The desired immunization process of antibody stimulation through vaccination was being profoundly prevented.
Three years later, at the National Institute for medical Research in London, virologists discovered similar causal effects on the growth if influenza virus in chicken egg membranes. Something was again naturally interfering with the growth of the virus after purposeful (unnatural) injection. In their research paper they coined this viral inhibitory factor as “Interferon“.
At the same time, back at the University of Tokyo, those same Japanese virologists finally discovered the essense of what they originally coined as “Viral Inhibitory Factor (VIF)”, and both research branches agreed that this anti-viral substance was caused by the same class of factors, and eventually these became officially known in medical science as “Interferon” (multiple types).
Further study revealed that these Interferon proteins reside in different human chromosomes, and a purification process of biologically active beta interferon was finally isolated in 1977. By the early 1980’s interferon protein types were isolated and cloned to show conclusive proof that indeed interferons were responsible for interfering with viral reproduction. Eventually, these interferon isolates were used as a treatment for viral infections.
So what are these naturally interfering produced factors, and why do pharmaceutical corporations hate them so much that they seek to interfere with their pre-programmed interference?
Clinically defined, Interferons (IFNs) are proteins (glycoproteins called cytokines) made and released by healthy host cells (naturally occurring cells in your body) in response to the presence of pathogens such as viruses, bacteria, parasites, or tumor cells. They literally act as communication devices traveling as RNA messengers, allowing cells to communicate with each other like micro text messages, creating a trigger effect to “interfere” with disease and viral replication by turning on the protective defensive structure of the immune system that is responsible for activating immune cells (natural killer cells, macrophages, etc.). Interferons also increase the ability of uninfected host cells in their ability to resist new infection by virus (an invading parasite to the host cell), and communicate the known presence of tumor cells to the immune system, up-regulating antigens to T lymphocytes.
A lymphocyte is one of 3 cells from the vertebrate’s immune system found in the lymphatic system called NK (natural killer) cells, B cells, or T cells. There are currently identified 10 distinct interferons (IFN’s), 7 of which are found in humans. These are further broken down by classes (types 1, 2, and 3). All of these IFN’s are vital for the body’s defense against disease states and infections as well as prevention of tumor growth.
In layman’s terms, we could say that all of the body’s naturally healthy cells send out cell-phone calls in the form of amino acids (proteins), which float through the body as if upon a wirelessly fluid Ethernet, directly connecting to the body’s receiving phone-line like a 911 emergency call; thus literally summoning the body’s first responders in the form of the immune system to send out little firefighter cells (Natural Killer, B, and T-cell lymphocytes) to stop the spread of the fire caused by viral, bacterial, parasitic, or tumor causing pathogens that are invading the host cells.
Those Pesky Little Interferons
While interferons have been used in some cases as a breakthrough yet totally underutilized treatment for the slowing or halting of certain disease growth in humans, we find a much more sinister reason for such research and identification of interferons in modern medicine and vaccine production. You see, the original discovery of interferons was not an altruistic attempt to isolate and synthesize an amino acid compound that would treat disease. In fact, far from it…
Back in 1956, those Japanese and British virologists were not trying to cure disease. No, they were trying to induce disease within their animal subjects for research purposes and spread it into chicken eggs so as to grow the disease for vaccination and “other” purposes; chicken embryo substrates being the most popular method for disease culture growth. But as they learned through continuous interference from the host subjects, something kept getting in the way of their purposeful disease infection of those hosts – an at the time unknown intracellular function of the body as of yet unknown, later to be named as Interferon.
Please understand… in order to vaccinate against disease, these scientists believed that they had to stop the bodies own natural defense against the very disease these scientists were trying to purposefully infect their test subjects with. Some might call this a paradox… or just insanity. In order for their pseudo-science to supposedly work, those virologists had to figure out a way to cut the cell-phone signaling process (now known as interferon) caused by their purposeful inoculate infection of the hosts. They needed to cause the body to cease in its perfectly natural capacity to fight the very disease they were injecting into it, so as to grow the disease within that host body. This would seem to the average person to be, on the surface and rightly so, a counter-productive effort on their part. But then the average person could never comprehend what was happening behind the scenes, let alone the true purpose of funding such experimental “science” as medicine.
Let’s take the phenomenon known as Auto-Immune Deficiency Syndrome (AIDS) for example…
What are its symptoms?
Rare cancerous tumors, viral-like infection, wasting syndrome, and general immune-supression of the lymphatic system.
Sound familiar? Like maybe the body’s phone-lines are down?
The body works though a system of communication devices in bilogical form. When one part or system of the body needs to communicate with another, it does so through a highly advanced structure of expressive signaling and transduction; the release of various types of cells, proteins, and other substances that trigger each inter-dependent system to respond in kind. It is this body-wide platform of cellular communication that is being attacked and blocked by the introduction of inhibiting factors like infectious prions and other melevolent substances.
The body works just fine until it is stung and thus injected (vaccinated) with foreign proteins, DNA, RNA, and other ingreedients that in no other way would ever be able to insert themselves into the body of man (or rabbit).
The main issue with AIDS patients is the lack of the body’s immune response regarding the production of T Lymphocytes, commonly called T-cells. For some reason, despite the body’s many dis-ease states as symptoms of the AID-syndrome, the body just isn’t getting the hint to produce the very thing that it needs to fight infection. It seems we have a failure to communicate here… For some reason the emergency 911 cell-phone lines seem to be cut, and the first responders (T-cells) are just not being called into action by the healthy cells that are under attack. Their chemical screams for help are going unheard. It’s as if the immune system labor union went on strike, and these “AIDS” symptoms are the resulting chaos and unrest that ensues throughout the body.
Not ironically, these are the same symptoms of what is known as Gulf War Syndrome, a known vaccine induced disease state thought by many researchers to be caused by vaccine adjuvants like squalene and other ingredients injected into the guinea pig soldiers of our military.
But what could possibly cause such a chain reaction throughout the body’s immune-supressive system?
What could possibly have been introduced within the body to prevent its ability to make a protein phone call, just like in those poor test-rabbits so many decades ago?
What is preventing interferon from interfering with the disease process, defeating its attempts to transmit its signal for help to the imune system?
Enter bioengineering and the novel prion…
So how could this novel disease state be simultaniusly spread
throughout Africa and eventually the first world?
Altering Gene Expression: Just A Little Pinprick
“The genetic code is universal….
The complete word-for-word universality of the genetic dictionary is,
for the taxonomist, too much of a good thing.”
–Evolutionist Richard Dawkins, in his book,
‘The Blind Watchmaker’ (1986, p. 270)
“It is recognized by molecular biologists that the genetic code is universal,
irrespective of how different living things are in their external appearances.”
–Creationist Robert Kautz, in his book,
‘The Origin of Living Things’ (1988, p. 44)
“The construction and metabolism of a cell are thus dependent
upon its internal ‘handwriting’ in the genetic code.
Everything, even life itself, is regulated from a biological viewpoint
by the information contained in this genetic code.
All syntheses are directed by this information.”
–A.E. Wilder-Smith, United Nations scientist (1976, p. 254).
“It may seem a platitude to say that the offspring of buttercups, sparrows and human beings are buttercups, sparrows and human beings… What then keeps them, and indeed living things in general, “on the right lines?” Why are there not pairs of sparrows, for instance, that beget robins, or some other species of bird: why indeed birds at all? Something must be handed on from parent to offspring which ensures conformity, not complete but in a high degree, and prevents such extreme departures. What is it, how does it work, what rules does it obey and why does it apparently allow only limited variation? Genetics is the science that endeavours to answer these questions, and much else besides. It is the study of organic inheritance and variation, if we must use more formal language.”
–British geneticist, E.B. Ford,
‘Understanding Genetics’ (1979, p. 13).
“Tablets of stone prepared by the Babylonians some 6,000 years ago have been interpreted as showing pedigrees of several successive generations of horses, thus suggesting a conscious effort toward improvement. Other stone carvings of the same period illustrate artificial cross-pollination of the date palm as practiced by the early Babylonians. The early Chinese, many years before the Christian era, improved varieties of rice. Maize was cultivated and improved in the western hemisphere by the American Indians, beginning at an early period in their history. In another era, Hippocrates, Aristotle, and other Greek philosophers made observations and speculations suggesting genetic principles.”
–Eldon Gardner, ‘The History of Biology’ (1972, pp. 399-400)
“And God said, let the earth put forth grass, the herb
yielding seed, and the fruit tree yielding fruit after its kind,
wherein is the seed thereof upon the earth, and it was so.
And the earth brought forth grass, the herb yielding seed and
the fruit tree yielding fruit after its kind whose seed was in itself.”
–The Bible, Genesis 1:11-12
“In the first chapter of Genesis, however, because it is a matter of the greatest religious importance, the Bible speaks clearly and finally on a matter of biology. After its kind is the statement of a biological principle that no human observation has ever known to fail. The most ancient human records engraved on stone or painted on the walls of caves bear witness to the fact that horses have ever been horses, bears have ever been bears, geese have ever been geese, reindeer have ever been reindeer. The most desperate and subtle efforts of man in modern times have been unable to alter this divine decree. The Bible teaches that from the beginning there have been a large number of types of living things, man included, which were so created as to remain true to their particular type throughout all generations…. The latest results of modern biological research, Mendel’s Laws, agree exactly with what was written by Moses three thousand years ago—and they also elucidate it…”
Byron Nelson, ‘After Its Kind’, (1967, pp. 3,103)
“…once a fertilized, (a) single human cell begins to develop, the original plans are
faithfully copied each time the cell divides (a process called mitosis)
so that every one of the thousand million million cells in my body, and in yours,
contains a perfect replica of the original plans for the whole body”.
–Evolutionist John Gribbin (1981, p. 193)
“The Nobel laureate, F.H. Crick has said that if one were to
translate the coded information on one human cell into book form,
one would require one thousand volumes each of five hundred pages to do so.
And yet the mechanism of a cell can copy faithfully at cell division
all this information of one thousand volumes each of
five hundred pages in just twenty minutes.”
–Dr. Wilder-Smith (1976, p. 258).
“Every organism has in it a store of what is called genetic information… I will refer to an organism’s genetic information store as its Library…. Where is the Library in such a multicellular organism? The answer is everywhere. With a few exceptions every cell in a multicellular organism has a complete set of all the books in the Library. As such an organism grows its cells multiply and in the process the complete central Library gets copied again and again…. The human Library has 46 of these cord-like books in it. They are called chromosomes. They are not all of the same size, but an average one has the equivalent of about 20,000 pages…. Man’s Library, for example, consists of a set of construction and service manuals that run to the equivalent of about a million book-pages together.”
“It is an indication of the sheer complexity of E. coli
that its Library runs to a thousand page-equivalent”
–A.G. Cairns-Smith (1985, pp. 9,10,11)
“The DNA in living cells contains coded information. It is not surprising that so many of the terms used in describing DNA and its functions are language terms. We speak of the genetic code. DNA is transcribed into RNA. RNA is translated into protein. Protein, in a sense, is coded in a foreign language from DNA. RNA could be said to be a dialect of DNA. Such designations are not simply convenient or just anthropomorphisms. They accurately describe the situation.”
–Lester and Bohlin (1984, pp. 85-86)
Further, consider that human beings have learned to store information on
clay tablets, stone, papyrus, paper, film, cassettes, microchips, etc.
Yet ‘human technology has not yet advanced to the point of
storing information chemically as it is in the DNA molecule‘
“It is not possible for a code, of any kind, to arise by chance or accident.
The laws of chance or probability have been worked out by mathematics…
A code is the work of an intelligent mind. Even the cleverest dog or chimpanzee
could not work out a code of any kind. It is obvious then that chance cannot do it…
This could no more have been the work of chance or accident than could the
“Moonlight Sonata” be played by mice running up and down the keyboard of my piano!
Codes do not arise from chaos”
–Professor Andrews (1978, pp. 28,29).
Infecting The World
So what happens when man comes clumsily and irresponsibly into the age of molecular science, where he begins to intermix species through inoculation? How can man know if his limits truly are what is written in the ancient scriptures and philosophies of moral men unless he seeks the answers by destroying the perfection of nature’s mathematical equations of the biology of life? How can we know the limits of genetically altered life if we don’t push those limits to the very brink of extinction of species, including our own?
Ancient warnings are for pussies!!!
In my previous research, I have postulated the horrifyingly evidence-based theory that all modern disease states, from the dementia’s to cancer to AIDS, have been induced through the vaccination process via the direct bodily injection of foreign “infectious” proteins called prions. Further research has all but confirmed the reality of this notion, showing that the inherent protective foundation of these cellular proteins in cell health (before infection) are essential to life itself.
Prion protein aids bone marrow
New study findings point to possible stem cell role for normal form of protein
The normal form of prion protein (PrP) appears necessary for bone marrow stem cells to renew themselves
, scientists reported online this week
in the Proceedings of the National Academy of Sciences
. These findings suggest a potential physiological function in stem cells for the normal form of the widely expressed protein. “Prior to this work there was no hint that PrP had a function in stem cell biology,” co-author Andrew Steele at the Whitehead Institute for Biomedical Research in Cambridge, Mass., told The Scientist
. “We are now looking into PrP function in other adult stem cells, particularly neural stem cells
.” Prions are infamous for being associated with transmissible spongiform encephalopathies (TSEs) such as mad cow disease, but the function of PrP — the normal, widespread and highly conserved form of prions — remains a mystery. In preliminary studies, co-author Cheng Cheng Zhang discovered 40% of adult mouse bone marrow cells expressed PrP on their surfaces
. More than 80% of these PrP-marked cells were red blood cells or their developmental precursors, suggesting PrP might be a marker for long-term hematopoietic stem cells, which can give rise to the entire adult blood system
. To determine if PrP was a marker for long-term hematopoietic stem cells, the researchers took bone marrow cells from wild-type mice and purified them into fractions, some of which expressed PrP. Six months after transplantation into lethally irradiated mice, the researchers saw both short- and long-term engraftment in mice that received PrP-containing cells, but only short-term engraftment activity in mice receiving non-PrP cells. While PrP is a marker for long-term hematopoietic stem cells in wild-type mice, PrP-knockout mice still possess these cells, as well as relatively normal levels of their derived progeny. To determine what function PrP might normally have in hematopoietic stem cells, the researchers carried out several rounds of bone marrow implantations
. First they transplanted bone marrow from either wild-type mice or a PrP-null strain into lethally irradiated mice. When the engrafted marrow flourished and generated peripheral blood cells, the researchers implanted the newly reconstituted bone marrow into another lethally irradiated mouse group, then repeated the process a third time
. In each round after the first, bone marrow originating from PrP-null mice experienced a dramatically reduced ability to renew itself, while cells from the wild-type mice did not
. Retroviral infections that expressed PrP in recipients of PrP-null bone marrow rescued this defective process, suggesting PrP is necessary for hematopoietic stem cell self-renewal
. Odile Kellerman at the Pasteur Institute in Paris, who did not participate in this study, noted prions often trigger neuron death in TSEs after long incubation periods
,” similarly, PrP only impacted hematopoietic stem cells over the long term
. “In both cases, PrP appears to contribute to the long-lasting adaptation of cells to injury
,” she told The Scientist
. Kellerman suggested that when PrP function is disrupted, cells try to adapt, “but in the long term, this turns out to be detrimental.” The exact mechanism behind how PrP might contribute to hematopoietic stem cell renewal remains unknown. Co-author Harvey Lodish speculated PrP might bond to and concentrate a hormone on the cell surface, or help stem cells adhere to neighboring cells or extracellular matrix
. “It should prove fairly straightforward to see if it is adhering to other proteins or any known or unknown hormones,” he told The Scientist
. William Stanford at the University of Toronto, who did not participate in this study, noted that PrP is tethered to cell membranes via a glycosylphosphatidylinositol (GPI) anchor, similar to hematopoietic stem cell marker Sca-1
. “This suggests these GPI-anchored proteins, which have similar functions, may operate through a common mechanism
,” Stanford told The Scientist
. Future experiments could investigate whether overexpressing PrP in hematopoietic stem cells increases self-renewal, and rescues self-renewal defects such as in the Sca-1 deficient mouse, Stanford added — or if genetically substituting PrP with a different GPI-anchored protein rescues the self-renewal defect seen in PrP-null mice. email@example.com Links within this article C.C. Zhang et al. “Prion protein is expressed on long-term repopulating hematopoietic stem cells and is important for their self-renewal
.” PNAS Early Edition.
Published online January 30, 2006. http://www.pnas.org
“Prion hypothesis proven?” The Scientist
, April 21, 2005. http://www.the-scientist.com/article/display/22653/
M. Fogarty. “Prions – The terminators.” The Scientist
, July 28, 2003. http://www.the-scientist.com/article/display/13974/
M. Fogarty. “Researchers further define sources of adult blood stem cells.” The Scientist
, September 16, 2002. http://www.the-scientist.com/article/display/13257/
J.U. Adams. “The tiniest of life’s rafts.” The Scientist
, October 11, 2004 http://www.the-scientist.com/article/display/14978/
Neurons and Astrocytes Respond to Prion Infection by Inducing Microglia Recruitment
The accumulation and activation of microglial cells at sites of amyloid prion deposits or plaques have been documented extensively. Here, we investigate the in vivo recruitment of microglial cells soon after intraocular injection of scrapie-infected cell homogenate (hgtsc+) using immunohistochemistry on retinal sections. A population of CD11b/CD45-positive microglia was specifically detected within the ganglion and internal plexiform retinal cell layers by 2 d after intravitreal injection of hgtsc+. Whereas no chemotactism properties were ascribed to hgtsc+ alone, a massive migration of microglial cells was observed by incubating primary cultured neurons and astrocytes with hgtsc+ in a time- and concentration-dependent manner. hgtsc+ triggered the recruitment of microglial cells by interacting with both neurons and astrocytes by upregulation of the expression levels of a broad spectrum of neuronal and glial chemokines. We show that, in vitro and in vivo, the microglia migration is at least partly under the control of chemokine receptor-5 (CCR-5) activation, because highly specific CCR-5 antagonist TAK-779 significantly reduced the migration rate of microglia. Activated microglia recruited in the vicinity of prion may, in turn, cause neuronal cell damage by inducing apoptosis. These findings provide insight into the understanding of the cell-cell communication that takes place during the development of prion diseases.
Prion hypothesis proven?
In vitro infectivity study in Cell stirs tempest in a test tube
April 21, 2005
Protein aggregates generated in a test tube infected wildtype hamsters with a disease much like scrapie, according to an article appearing this week in Cell. Such a demonstration has, in the past, been called the gold standard of proof for the prion hypothesis, Stanley Prusiner’s Nobel-winning assertion that infectious, self-replicating protein isoforms are the culprit in transmissible spongiform encephalopathies (TSEs) like scrapie, Creutzfeldt-Jakob disease, and mad cow disease.
Study coauthor Claudio Soto, said that this demonstration, together with a paper published by Prusiner’s group last summer, should allay most doubts. “There is really little room for skepticism,” he told The Scientist.
But the study has done little to quiet prion hypothesis skeptics. “I’m not going to abandon alternative hypotheses for the time being,” said Robert A. Somerville of the Institute for Animal Health, Edinburgh.
While Prusiner’s group had successfully infected a mouse with a recombinant protein derived from bacteria, some argued that their use of transgenic mice susceptible to the disease undercut the power of the demonstration. In the new study, researchers at the University of Texas Medical Branch, Galveston, Universidad Autonoma, Madrid, and the University of Chile in Santiago fine-tuned a cyclical process for amplifying aggregated protein from an infected hamster brain. Through serial dilutions, they were able to infect a wildtype hamster with in vitro–produced aggregates without any traces of the original infectious brain. But skeptics, including a member of Prusiner’s group, argue that using material from a diseased hamster brain could have resulted in residual contamination.
Soto’s group has been using a process that they call protein misfolding cyclic amplification (PMCA), which aids the aggregation of the normal cellular protein PrPc into the misfolded, polymer-forming PrPres that is associated with TSE pathology. The process works in a fashion similar to polymerase chain reaction (PCR) amplification of oligonucleotides. After seeding PrPc with PrPres, the solution is incubated and sonicated. “Once the aggregates become long enough, we split them into smaller pieces so that in a new conversion, a new incubation, they are able to convert more and more of the normal protein,” Soto explained.
Crucially, however, the PrPres “seed” comes from infected hamster brain homogenate, while the normal PrPc comes from healthy hamster brain homogenate. “They actually started from infectious material, and we didn’t,” said Giuseppe Legname, of the University of California, San Francisco, and co-author on the Prusiner paper. “It’s an alternative approach to demonstrate that you might make prions, but to say that these are synthetic prions, it’s very difficult.”
Soto insisted that serial dilutions between rounds of PMCA reduce scrapie brain homogenate to an amount equivalent to a 10 to the minus 10th and a 10 to the minus 20th–fold dilution. Infectivity generally drops off after 10 to the minus 9th, according to the paper. “We’ve completely ruled out the possibility that the infectivity is still remaining from… the original brain,” Soto said...
While the article continues to criticize the control group results, which you may read at the link above, the important point here is that scientists are creating prions and making them purposefully more infectious. They are testing them in various substances and frequencies. And through the ultra-sound sonic vibration described above as protein misfolding cyclic amplification (PMCA), they are able to excite the growth factor of infectious prions so that they take over (mis-fold) healthy brain tissue much quicker. This PMCA process is used in autopsy to detect prion disease.
I have my own concerns that these ultra-sound frequencies are the same as used in cell-phone towers and in the process of ultra sound for unborn infants and other medical procedures, as well as other frequencies unknown via smart meters, radio waves, etc. We are playing with the fuel for the fire and there is virtually no escaping this permanent state of sonic bombardment…
It is also interesting to note that two men wsere cured of AIDS symptoms by receiving a bone marrow transfusion not so long ago…
(CBS News) Two men who’ve had HIV for years may now be free of the disease following bone marrow transplants, researchers at Brigham and Women’s Hospital in Boston announced Thursday.
The new research has some attendees at the XIX International AIDS Conference in Washington, D.C. hopeful for a cure.
Timothy Ray Brown, man thought to be first “cured” of AIDS, says he’s still cured
Man “cured” of AIDS: Timothy Ray Brown
Both patients were being treated for cases of cancer. One of the patients underwent a bone marrow transplant two years ago at the Dana-Farber/Brigham and Women’s Cancer Center in Boston, the other had the procedure done four years ago at the same hospital. NBCNews.com reports that one of the patients is in his 50s and has been infected since the early 1980s towards the beginning of the AIDS epidemic and the other man, in his 20s, was infected at birth.
Both stayed on their antiretroviral medication regimens, the standard treatment of HIV, following the transplants.
The researchers discovered that overtime as the patients’ cells were replaced by cells from the donor, evidence of HIV in the patients’ blood tests disappeared. The researchers also said both patients have no signs of HIV in their DNA or RNA and levels of their disease-fighting antibodies have also decreased. The researchers think the medications helped allow these cells to be replaced.
“This gives us some important information,” one of the researchers Dr. Daniel Kuritzkes, an infectious disease specialist at the hospital and Harvard Medical school said in a press release. “It suggests that under the cover of antiretroviral therapy, the cells that repopulated the patient’s immune system appear to be protected from becoming re-infected with HIV.”
The researchers themselves won’t call it a cure yet, saying they still need to check more tissues for traces of the disease. But they were surprised to see no signs of HIV beyond what’s seen in a blood test.
“We expected HIV to vanish from the patients’ plasma, but it is surprising that we can’t find any traces of HIV in their cells,” said co-resarcher Dr. Timothy Henrich, also of BWH and Harvard. “The next step is to determine if there are any traces of HIV in their tissue.”
The researchers’ announcement comes days after Timothy Ray Brown, the man known as the “Berlin Patient,” held a press conference in Washington, D.C., to say he’s still cured of AIDS five years after undergoing a bone marrow blood transplant…
It is important to note that the chemokine receptor-5 (CCR-5) antagonist prevents the cellular binding of the HIV-1 virus, as is explained in this video:
And how do these prions effect disease states?
Let’s take for example Multiple Sclerosis:
“The etiology of Multiple Sclerosis (MS) is unknown. Existing epidemiologic data suggests that MS can be an infectious disease. MS used to be classified as one of the ‘slow infections‘–many of these are caused by prions. Prions are small, proteinaceous, infectious particles–distinguished from viruses by the absence of intrinsic nucleic acids. In a contrast to the ‘classic’ prional diseases (Kuru, Scrapie or Creutzfeldt-Jacob Disease) that in CNS affect primarily neurons, the ‘target’ cell in MS is an oligodendrocyte. This may explain differences in disease presentation. This paper presents a pathophysiological model of MS based on the assumption that MS is a prional disease. Processes leading to the demyelination in Multiple Sclerosis seem also to involve lymphocytes, astrocytes and macrophages as well as the interferon system…”
NOTE: The protein that prions are made of (PrP) is found throughout the body, even in healthy people and animals, and necessarily protects cells from infections. However, PrP found in infectious material has a different structure and is resistant to proteases, the enzymes (proteins) in the body that can normally break down other proteins. The normal form of the protein is called PrPC, while the infectious form is called PrPSc — the C refers to healthy ‘cellular‘ PrP, while the Sc refers to infectious ‘scrapie‘, the prototypic prion disease, occurring in sheep. The infectious isoform of PrP, known as PrPSc, is able to convert normal PrPC proteins in humans into this infectious isoform by changing their conformation, or shape. This, in turn, alters the way the proteins interconnect, creating symptoms like transmissible spongiform encephalopathy (holes in the human brain like mad cow disease). PrPSc always causes prion disease. In the end, no cellphone call can be made if the interferon protein is infected and mis-folded before it is able to reach its receiving protien that would activate T-Cells or other immune responses. Another word for mis-fold might be easier to understand as to misinform. The immune system is being lied to in a strange, chemically unbalanced way due to prion protein infections (mis-folding). Sheep blood (serum) is a popular vaccine substrate to grow vaccines for humans upon, and the protein and DNA cannot be filtered out of the final vaccine product. There are no other viable explanations why infectious prions from animals would intermingle within a human body (xenotransplantaion/xenografting).
A thorough and sourced description about prions can be found here: http://www.omim.org/entry/176640
This interference that infectious prions cause to interferon and other protein-based signaling and transcription cells is shown in the research studies below. For those with the gumption, let’s play a biological game of connect the dots.
Prion infection is accelerated in (interferon type 3) IRF3-deficient mice…
The IRF3-dependent pathway is protective against prion infection in cell culture.
We tested whether over-expression of IRF3 (interferon) could affect the production of PrPSc (infectious/mis-folded prions) in the cell culture models. The level of PrPC (healthy prions) was not affected by the transient expression of the genes in uninfected N2a58 cells (data not shown). PrPSc was significantly decreased by overexpression of IRF3 in the 22L-N2a58 cells (A). We confirmed that the activated form of IRF3 (phosphorylated at Ser396 of IRF3) increases in a dose-dependent manner after transfection of the IRF3 gene in both 22L-N2a58 cells (A) and uninfected N2a58 cells (data not shown), indicating that the upregulation of IRF3 phosphorylation seen in the A is most likely due to an increase in the level of IRF3 protein after transfection.
To investigate the effect of downregulation of IRF3 in the 22L-N2a58 cells, we performed knockdown experiments using small interfering RNAs (siRNAs). IRF3 expression was significantly decreased by two types of siRNAs against IRF3, whereas β-actin expression, as the internal standard, was not changed (B)… These data suggest that IRF3 has an inhibitory effect on the production of PrPSc in the 22L-N2a58 cells.
To further evaluate the protective effect of IRF3… After incubation with 22L-infected BH (22L-BH), the cell clones were subcultured for five passages and analyzed by Western blotting with anti-PrP antibodies. The values of the PrPSc/PrPC ratio were inversely correlated with the values of the IRF3/beta-actin ratio (C), indicating that enhanced expression of IRF3 effectively blocks new prion infection.
In the present study, we found that a genetic deficiency of IRF3 accelerates the progression of TSE (transmissable prion disease) following i.p. transmission in mice and that the accumulation rate of PrPSc in the spleen is increased in the IRF3−/− mice. Furthermore, we demonstrated that IRF3 has an inhibitory effect on PrPSc accumulation and that the levels of IRF3 are inversely correlated with resistance to prion infection in cell culture.
IRF3 is known to be constitutively expressed in many tissues and cells (6, 22, 45). Indeed, we confirmed the expression of IRF3 in brains (data not shown) and N2a58 cells (). Furthermore, not only glial cells but also neurons express most innate immunity-related genes and produce type I IFN in response to virus infection (11). Although the role of IRF3 in prion propagation into the CNS is still unclear, we speculate that an absence of IRF3 signaling leads to increased prion replication not only in peripheral tissues but also in the CNS. It would be of great value to examine this further using neuron-specific IRF3-disrupted mice or neuron-specific IRF3-expressing mice.
It was reported in prion infection that genetic disturbance of TLR4 (36) or interleukin-10 (IL-10) (41) leads to shorter incubation periods of prion infection. Since these, respectively, are an upstream and a downstream factor of the IRF3-mediated pathway, the findings may be due in part to functional changes in IRF3-mediated signaling.
Based on these results, two hypothetical models are proposed to explain the inhibitory effect of IRF3 on the prion infection. The first is that MyD88-independent pattern recognition receptors (PRRs), such as TLR3, TLR4, or RIG-I/MDA5, might recognize prion, and the resulting activation of IRF3 could induce various IRF3-responsive genes that may participate in the protective effect. The fact that the in vivo administration of IFNs (interferons), a representative of the IRF3-responsive genes, previously failed to show inhibitory effects on TSE (13, 16) suggests that IRF3-responsive genes other than IFNs may be important for the inhibitory effect of IRF3 on prion infection. Of note, the protective effect of IRF3 against several viruses has been suggested to be largely independent of the production of type I IFN and is probably responsible for the antiviral actions of specific IRF3-responsive genes (10, 18, 21). Peritoneal macrophages from wild-type mice moderately induced tumor necrosis factor alpha (TNF-α) or IL-6 following exposure to PrPSc-mimicking PrP peptides (PrP residues 106 to 126 or PrP residues 118 to 135), whereas TLR4 signaling-mutant mice were impaired in their ability to produce these cytokines (36), supporting in part the hypothesis that some PRRs may sense PrPSc as a sort of PAMP. On the other hand, it should be noted that the MyD88-independent pathway activates both NF-κB and IRF3. Although the induction of proinflammatory cytokines essentially depends upon NF-κB, it was unclear whether the activation of IRF3 was induced by these PrP peptides. In fact, the hallmarks of IRF3 activation, such as phosphorylation, dimerization, and cytoplasm-to-nucleus translocation of IRF3 in 22L-N2a58 cells, were not detected (data not shown). Moreover, it was previously reported that IFNs were not detected in the serum, spleens, or brains of mice infected with scrapie (44). In addition, IFN-β mRNA does not increase in the brains of CJD (human prion disease) patients (7) or mice infected with ME7 prion strain (14). Hence, these results argue against the notion that the IRF3-mediated signaling is activated by prion infection, but it remains to be determined whether transient and weak responses are evoked at an early phase in the infection. The question as to whether IRF3-mediated signaling directly suppresses the production of PrPSc or increases its degradation also remains open.
Another explanation is that prion infection itself may have little effect on the pathway but that the basal activity of IRF3 may have some degree of inhibitory effect on prion propagation. It has been reported that IRF3 can be activated not only by viruses but also by multiple activators such as cellular stress and DNA damage (24, 34). Accordingly, it is possible that constitutive activation of IRF3, albeit at a low level, occurs in the brain even in the absence of a pathogen. This notion is further supported by the fact that constitutive, weak IFN signaling in the absence of viral infection plays a role in modifying cellular responsiveness in the immune and other biological systems (38, 40). Accumulating evidence indicates that many viruses have evolved to evade the innate immune system, including IRF3-mediated signaling (15, 23). For instance, an active mutant of IRF3 has been reported to exert a markedly suppressive effect on cellular HIV-1 infection, and administration of poly(I·C) potently inhibits HIV-1 replication in microglia through a pathway requiring IRF3. Nonetheless, HIV-1 itself does not activate IRF3 but, rather, decreases IRF3 protein in HIV-1-infected cells (12, 37). Likewise, prion infection might disturb the activation of IRF3 even though prion is considered to be largely composed of PrPSc. We are currently investigating this possibility. Furthermore, an analogy can be made between the role of IRF3 in prion infection and that of IL-10. The levels of IL-10 are not increased in the brains of scrapie-infected mice (14, 42), whereas IL-10 knockout mice are highly susceptible to the development of scrapie (41).
In conclusion, we have shown that IRF3, a key transcription factor of the MyD88-independent pathways, operates in the host defense machinery against prion infection. The findings provide new insight into understanding of the innate immunity to prion infection.
Interleukin-10 (IL-10), also known as human cytokine synthesis inhibitory factor (CSIF), is an anti-inflammatory cytokine. In humans, IL-10 is encoded by the IL10 gene.
Gene and protein structure
The IL-10 protein is a homodimer; each of its subunits is 178-amino-acid long.
IL-10 is classified as a class-2 cytokine, a set of cytokines including IL-19, IL-20, IL-22, IL-24 (Mda-7), and IL-26, interferons (IFN-alpha, -beta, -epsilon, -kappa, -omega, -delta, -tau, and -gamma) and interferon-like molecules (limitin, IL-28A, IL-28B, and IL-29).
Expression and synthesis
In humans, IL-10 is encoded by the IL10 gene, which is located on chromosome 1 and comprises 5 exons, and is primarily produced by monocytes and, to a lesser extent, lymphocytes, namely type 2 T helper cells (TH2), mastocytes, CD4+CD25+Foxp3+ regulatory T cells, and in a certain subset of activated T cells and B cells.
In biochemistry, a dimer is a macromolecular complex formed by two, usually non-covalently bound, macromolocules like proteins or nucleic acids. It is a quaternary structure of a protein.
A homo-dimer would be formed by two identical molocules (a process called homodimerization). A hetero-dimer would be formed by two different macromolecules (called heterodimerization).
Most dimers in biochemistry are not connected by covalent bonds. An example of a non-covalent heterodimer would be the enzyme reverse transcriptase, which is composed of two different amino acid chains. An exception is dimers that are linked by disulfide bridges such as the homodimeric protein NEMO.
Some proteins contain specialized domains to ensure dimerization (dimerization domains).
Examples of Homodimer include anti-bodies and Factor VII.
Microglia are a type of glial cell that are the resident macrophages of the brain and spinal chord, and thus act as the first and main form of active immune defense in the central nervous system (CNS).
Microglia constitute 10-15% of the total glial cell population within the brain. Microglia (and astrocytes) are distributed in large non-overlapping regions throughout the brain and spinal cord. Microglia are constantly scavenging the CNS for plaques, damaged neurons and infectious agents. The brain and spinal cord are considered “immune privileged” organs in that they are separated from the rest of the body by a series of endothelial cells known as the blood-brain barrier, which prevents most infections from reaching the vulnerable nervous tissue. In the case where infectious agents are directly introduced to the brain or cross the blood–brain barrier, microglial cells must react quickly to decrease inflammation and destroy the infectious agents before they damage the sensitive neural tissue. Due to the unavailability of antibodies from the rest of the body (few antibodies are small enough to cross the blood brain barrier), microglia must be able to recognize foreign bodies, swallow them, and act as antigen-presenting cells activating T-cells. Since this process must be done quickly to prevent potentially fatal damage, microglia are extremely sensitive to even small pathological changes in the CNS. They achieve this sensitivity in part by having unique potassium channels that respond to even small changes in extracellular potassium.
Microglial cells differentiate in the bone marrow from hematopoietic stem cells, the progenitors of all blood cells. During hematopoiesis, some of these stem cells differentiate into monocytes and travel from the bone marrow to the brain, where they settle and further differentiate into microglia.
Monocytes can also differentiate into myeloid dendritic cells and macrophages in the peripheral systems. Like macrophages in the rest of the body, microglia use phagocytic and cytotoxic mechanisms to destroy foreign materials. Microglia and macrophagesboth contribute to the immune response by acting as antigen presenting cells, as well as promoting inflammation and homeostatic mechanisms within the body by secreting cytokines and other signaling molecules.
In their downregulated form, microglia lack the MHC class I/MHC class II proteins, IFN-γ cytokines, CD45 antigens, and many other surface receptors required to act in the antigen-presenting, phagocytic, and cytotoxic roles that hallmark normal macrophages. Microglia also differ from macrophages in that they are much more tightly regulated spatially and temporally in order to maintain a precise immune response.
Another difference between microglia and other cells that differentiate from myeloid progenitor cells is the turnover rate. Macrophages and dendritic cells are constantly being used up and replaced by myeloid progenitor cells which differentiate into the needed type. Due to the blood brain barrier, it would be fairly difficult for the body to constantly replace microglia. Therefore, instead of constantly being replaced with myeloid progenitor cells, the microglia maintain their status quo while in their quiescent state, and then, when they are activated, they rapidly proliferate in order to keep their numbers up. Bone chimera studies have shown, however, that in cases of extreme infection the blood-brain barrier will weaken, and microglia will be replaced with haematogenous, cart-marrow derived cells, namely myeloid progenitor cells and macrophages. Once the infection has decreased the disconnect between peripheral and central systems is reestablished and only microglia are present for the recovery and regrowth period.
Transport of prion protein across the blood–brain barrier
The cellular form of the prion protein (PrPc) is necessary for the development of prion diseases and is a highly conserved protein that may play a role in neuroprotection. PrPc is found in both blood and cerebrospinal fluid and is likely produced by both peripheral tissues and the central nervous system (CNS). Exchange of PrPc between the brain and peripheral tissues could have important pathophysiologic and therapeutic implications, but it is unknown whether PrPc can cross the blood–brain barrier (BBB). Here, we found that radioactively labeled PrPc crossed the BBB in both the brain-to-blood and blood-to-brain directions. PrPc was enzymatically stable in blood and in brain, was cleared by liver and kidney, and was sequestered by spleen and the cervical lymph nodes. Circulating PrPc entered all regions of the CNS, but uptake by the lumbar and cervical spinal cord, hypothalamus, thalamus, and striatum was particularly high. These results show that PrPc has bidirectional, saturable transport across the BBB and selectively targets some CNS regions. Such transport may play a role in PrPc function and prion replication.
Cellular prion protein (PrPc) is perhaps best known as a source for the misfolded protein PrPsc (Prusiner, 1997) and as a prerequisite for the development of prion diseases (Mallucci et al., 2000). However, PrPc itself likely has important biological functions. It is found circulating in blood (Volkel et al., 2001) and is found in even higher levels in the cerebrospinal fluid (CSF) (Picard-Hagen et al., 2006). After ischemic events, PrPc levels increase in blood (Mitsios et al., 2007) and in neurons and brain endothelial cells in the peri-infarct region (Mitsios et al., 2007; Weise et al., 2004). These increases may reflect cytoprotective and neuroprotective roles for PrPc as recently reviewed (Roucou & LeBlanc, 2005). PrPc null mice have larger infarct volumes after ischemic events (Weise et al., 2006; Nasu-Nishimura et al., 2008) and more neuronal apoptosis after viral infections (Nasu-Nishimura et al., 2008) than wild type mice. In comparison, mice that overexpress PrPc have smaller infarcts and better neurological outcomes than wild type mice after ischemic events (Shyu et al., 2005). These protective events are likely mediated by PrPc through activation of anti-apoptotic (Spudich et al., 2005) and anti-oxidant pathways (White et al., 1999).
Sources of circulating PrPc likely include platelets (Robertson et al., 2006), endothelial cells (Simak et al., 2002), and lymphocytes (Politopoulou et al., 2000). Among lymphocytes, CD3 and CD8 lymphocytes have especially high levels which increase with aging (Politopoulou et al., 2000). All these cells have membrane bound PrPc that apparently can be released into the circulation. Platelet activation (Robertson et al., 2006) or endothelial apoptosis (Simak et al., 2002), for example, results in release of PrPc from those cells.
Thus, PrPc occurs in both blood and in CSF with levels that are likely responsive to disease states. This raises the question of whether PrPc can cross the blood–brain barrier (BBB). Such passage could link the two pools of PrPc and the events that control their levels. Here, we examined the ability of PrPc to cross the BBB in both the blood-to-brain and the brain-to-blood directions.
Capillary depletion as modified for use in the mouse (Triguero et al., 1990; Gutierrez et al., 1993) was used to determine the degree to which PrPc was sequestered and retained by the vascular bed of the brain.
I-PrPc was also taken up by the peripheral tissues of spleen, liver, kidney and cervical lymph nodes ()… there was a statistically significant decrease in the Ki for brain: F(1,8) = 7.97, p <0.05. This demonstrates that transport of PrPc across the BBB involves a saturable transport system.
shows values for brain and spinal cord regions. Statistical comparison of the whole brain value to brain regions and olfactory bulb (spinal cord regions excluded) showed a statistically significant variation: F(22,62) = 18.3, p <0.001. The hypothalamus, thalamus, and striatum showed statistically (p <0.01) greater uptake in comparison to whole brain. The highest uptake, however, was into the lumbar region of the spinal cord. Inhibition of uptake by unlabeled PrPc (; p <0.05) was found for whole brain, olfactory bulb, 4 of the 10 brain regions (occipital cortex, thalamus, striatum, and midbrain) and two of the spinal cord regions (cervical and lumbar)…
Brain-to-blood efflux of PrPc after icv injection. Half-time clearance from brain was 15.7 min. Inset shows that inclusion of unlabeled PrPc in the icv injection increased retention of radioactively labeled PrPc by brain, demonstrating a saturable component…
Does aluminum in vaccines have a more sinister plot that is stated?
Differential effect of aluminum on the blood-brain barrier transport of peptides, technetium and albumin.
Aluminum is a neurotoxin capable of altering membrane structure and function. We investigated whether aluminum also can affect saturable transport across membranes using the blood-brain barrier as our model. Mice were given i.p. or i.v. aluminum (up to 100 mg/kg) as the chloride salt and the disappearance from the brain of several centrally administered substances was measured. We found that aluminum rapidly and profoundly inhibited the saturable system that transports the small, N-tyrosinated peptides Tyr-MIF-1 and the enkephalins from the brain to the blood by acting as a noncompetitive inhibitor. In contrast, the disappearance from the brain of technetium pertechnetate (a substance also transported out of the brain by a different saturable system), albumin or D-Tyr-MIF-1 (a stereoisomer of Tyr-MIF-1 that was confirmed not to be transported by the carrier system) was not affected by aluminum. Aluminum also did not alter either the saturable or nonsaturable component of the uptake of Tyr-MIF-1 by erythrocytes. These findings suggest that one mechanism by which aluminum may induce neurotoxicity is by selective alteration of the transport systems of the blood-brain barrier.
An enkephalin is a pentapeptide involved in regulating nociception in the body. The enkephalins are termed endogenous ligands, as they are internally derived and bind to the body’s opioid receptors. Discovered in 1975, two forms of enkephalin were revealed, one containing leucine (“leu”), and the other containing mathione (“met”). Both are products of the proenkephalin gene.
Endogenous opioid peptides
There are three well-characterized families of opioid peptides produced by the body: enkephalins, endorphines, and dynorphins. The met-enkephalin peptide sequence is coded for by the enkephalin gene; the leu-enkephalin peptide sequence is coded for by both the enkephalin gene and the dynorphin gene. The proopiomelanocortin gene (POMC) also contains the met-enkephalin sequence on the N-terminus of beta-endorphin, but the endorphin peptide is not processed into enkephalin.
Main article: Opioid recepter
The receptors for enkephalin are the delta opioid receptors. Opioid receptors are a group of G-protein-coupled receptors, with other opioids as ligands as well. The other endogenous opioids are dynorphins (that bind to kappa receptors), endorphines (mu receptors), endomorphins, and nociceptin/orphanin FQ. The opioid receptors are ~40% identical to somatostatin receptors (SSTRs).
Endomorphins, Met-Enkephalin, Tyr-MIF-1, and the P-glycoprotein Efflux System
The P-glycoprotein (P-gp) transport system, responsible for the efflux of many therapeutic drugs out of the brain, recently has been shown to transport the endogenous brain opiate endorphin. We used P-gp knockout mice (Mdr1a) and their controls to determine where P-gp is involved in the saturable efflux systems of four other endogenous opiate-modulating peptides across the blood-brain barrier (BBB). After injection of endomorphin-1 (Tyr-Pro-Trp-Phe-NH2), endomorphin-2 (Tyr-Pro-Phe-Phe-NH2), Met-enkephalin (Tyr-Gly-Gly-Phe-Met-OH), and Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH2) into the lateral ventricle of the mouse brain, residual radioactivity was measured at 0, 2, 5, 10, and 20 min later. The results showed no difference in the disappearance of any of these peptides from the brains of knockout mice compared with their controls. This demonstrates that unlike endorphin and morphine, P-gp does not seem to be required for the brain-to-blood transport of the endomorphins, Met-enkephalin, or Tyr-MIF-1 across the BBB.
Endorphins (“endogenous morphine”) are endogenous opioid inhibitory neuropeptides. They are produced by the central nervous system and pituitary gland. The term implies a pharmacological activity (analogous to the activity of the corticosteroid category of biochemicals) as opposed to a specific chemical formulation. It consists of two parts: endo- and -orphin; these are short forms of the words endogenous and morphine, intended to mean “a morphine-like substance originating from within the body.”
Opioid neuropeptides were first discovered in 1974 by two independent groups of investigators:
- John Hughes and Hans Kosterlitz of Scotland isolated — from the brain of a pig — what some called enkephalins (from the Greek εγκέφαλος, cerebrum).
- Around the same time, in a calf brain, Rabi Simantov and Solomon H. Snyder of the United States found what Eric Simon (who independently discovered opioid receptors in vertebral brains) later termed “endorphin” by an abreviation of of “endogenous morphine”, meaning “morphine produced naturally in the body”. Importantly, recent studies have demonstrated that human and diverse animal tissues are in fact capable of producing morphine itself, which is not a peptide.
Mechanism of action
Beta-endorphin (β-endorphin) is released into blood from the pituitary gland and into the spinal cord and brain from hypothalamic neurons. The β-endorphin that is released into the blood cannot enter the brain in large quantities because of the blood-brain barrier, so the physiological importance of the β-endorphin that can be measured in the blood is far from clear. β-endorphin is a cleavage product of pro-opiomelanocortin (POMC), which is also the precursor hormone for adrenocorticotrophic hormone (ACTH). The behavioural effects of β-endorphin is exerted by its actions in the brain and spinal cord, and it is presumed that the hypothalamic neurons are the major source of β-endorphin at those sites. In situations where the level of ACTH is increased (e.g., Cushing’s disease), the level of β-endorphin also increases slightly.
β-endorphin has the highest affinity for the μ1 opioid receptor, slightly lower affinity for the μ2 and δ opioid receptors, and low affinity for the κ1 opioid receptors. μ-Opioid receptors are the main receptor through which morphine acts. In the classical sense, μ opioid receptors are presynaptic, and inhibit neurotransmitter release. Through that mechanism, they inhibit the release of the inhibitory neurotransmitter GABA, and disinhibit the dopamine pathways, causing more dopamine to be released. By hijacking this process, exogenous opioids cause inappropriate dopamine release, and can lead to aberrant synaptic plasticity, which can cause dependency. Opioid receptors have many other and more important roles in the brain and periphery; however, modulating pain, cardiac, gastric and vascular function as well as possibly panic and satiation. Also, receptors are often found at postsynaptic locations as well as at presynaptic locations…
Morphine preconditioning reduces lipopolysaccharide and interferon-γ-induced mouse microglial cell injury via δ1 opioid receptor activation
Microglial cells play an important role in the inflammatory response of a broad range of brain diseases including stroke, brain infection and neurodegenerative diseases. However, there is very little information regarding how to protect microglial cells. Here, we showed that incubation of the C8-B4 mouse microglial cells with lipopolysaccharide (LPS) plus interferon-γ (IFNγ) induced cytotoxicity as assessed by the amount of lactate dehydrogenase (LDH) released from the cells. Preconditioning the cells with morphine for 30 min concentration-dependently reduced LPS plus IFNγ-induced cell injury. This morphine preconditioning effect was abolished by naloxone, a general opioid receptor antagonist, by naltrindole, a selective δ opioid receptor antagonist and by 7-benzylidenenaltrexone maleate, a selective δ1 opioid receptor antagonist. However, this protective effect was not affected by β-funaltrexamine, a selective μ opioid receptor antagonist, nor-binaltorphimine, a selective κ opioid receptor antagonist or naltriben, a selective δ2 opioid receptor antagonist. LPS plus IFNγ induced the expression of inducible nitric oxide synthase (iNOS), which was not affected by morphine preconditioning. Our results suggest that morphine induced a preconditioning effect in microglial cells. This effect may be mediated by δ1 opioid receptors and may not be through inhibiting the expression of iNOS, a potentially harmful protein.
Prion peptide PrP106-126 induces inducible nitric oxide synthase (iNOS) and proinflammatory cytokine gene expression through the activation of NF-kB in macrophage cells
The inflammatory response in prion diseases is dominated by microglia activation. The molecular mechanisms that lie behind this inflammatory process are not very well understood. In the present study, we examined the activation of nuclear factor-kappa B (NF-κB) upon exposure to PrP106-126 and its role in PrP106-126-induced upregulation of inducible nitric oxide synthase (iNOS) and proinflammatory cytokines (interleukin [IL]-1β, tumor necrosis factor [TNF]-α, IL-6) in Ana-1 macrophages. The results showed that iNOS and proinflammatory cytokine release was significantly elevated in Ana-1 macrophages upon exposure to PrP106-126; that PrP106-126 treatment led to a significant NF-κB activation; that proinflammatory cytokines gene expression was elevated in macrophages upon exposure to PrP106-126; and that NF-κB inhibition significantly abrogated PrP106-126-induced upregulation of iNOS and inflammatory cytokine mRNA expression. These results suggest that treatment with neurotoxic prion peptides leads to the activation of transcription factor NF-κB, which in turn stimulates gene expression of iNOS and proinflammatory cytokines in Ana-1 macrophages.
The Transcription Factor Nuclear Factor-kappa B and Cancer
Since the discovery of nuclear factor-kappa B (NF-κB) in 1986, many studies have been conducted showing the link between the NF-κB signalling pathway and control of the inflammatory response. Today it is well known that control of the inflammatory response and apoptosis is closely related to the activation of NF-κB. Three NF-κB activation pathways exist. The first (the classical pathway) is normally triggered in response to microbial and viral infections or exposure to pro-inflammatory cytokines that activate the tripartite IKK complex, leading to phosphorylation-induced IκB degradation and depends mainly on IKKβ activity. The second (the alternative pathway), leads to selective activation of p52:RelB dimers by inducing the processing of the NF-κB2/p100 precursor protein, which mostly occurs as a heterodimer with RelB in the cytoplasm. This pathway is triggered by certain members of the tumour necrosis factor cytokine family, through selective activation of IKKα homodimers by the upstream kinase NIK. The third pathway is named CK2 and is IKK independent. NF-κB acts through the transcription of anti-apoptotic proteins, leading to increased proliferation of cells and tumour growth. It is also known that some drugs act directly in the inhibition of NF-κB, thus producing regulation of apoptosis; some examples are aspirin and corticosteroids. Here we review the role of NF-κB in the control of apoptosis, its link to oncogenesis, the evidence of several studies that show that NF-κB activation is closely related to different cancers, and finally the potential target of NF-κB as cancer therapy.